Abstract

In this article, protocols are described for preparation of digoxigenin-labeled oligo-DNA probe and their use in in situ hybridization. As has been shown previously (Koji et al. 1988, 1989; Koji and Brenner 1993; Koji and Nakane 1990, 1996), nonradioactive synthetic oligo-DNA probes can be very useful tools. The 3’ -end of oligo-DNA can be labeled enzymaticallywith several possible haptens, such as digoxigenin and biotin. Digoxigenin, a plant steroidal aglycone, is especially suitable for immunocytochemical detection because its epitope is not present in animal cells. Also, it has recently been confirmed that under appropriate conditions, there is no substantial difference in the sensitivity of detection between [35S] cRNA (Denjin et al. 1990) or [35S] cDNA (Unger et al. 1991) probes and nonradioactive probes.